219株阴沟肠杆菌对16种抗菌药物耐药性的研究

目的：了解阴沟肠杆菌的耐药状况;方法：测定近三年分离的２１９株阴沟肠杆菌对１６种抗生素的耐药率;结果：阴沟肠杆菌具有较高的耐药率及多重耐药性且有耐药率逐年上升趋势,其机制可能与细菌产生β内酰胺酶、外膜微孔蛋白改变及青霉素结合蛋白改变三方面有关     

229株阴沟肠杆菌的标本分布及耐药性分析

目的 了解阴沟肠杆菌在医院感染的标本分布和耐药情况,为临床合理选择和应用抗生素提供依据.方法 采用VITEK 60全自动微生物分析仪和配套的GNI、GNS-143、GNI-448,对229株阴沟肠杆菌进行分离鉴定和药敏试验,药敏结果使用WHONET 5.5软件进行分析.结果 从2007年1月至2011年9月共分离到阴沟肠杆菌229株,59.0％来自于呼吸道标本,其次是尿液占13.5％,再次为创面分泌物/脓液占12.7％.阴沟肠杆菌对美洛培南、亚胺培南和头孢哌酮/舒巴坦的耐药率分别为0.0％、0.4％和2.5％,对氨苄西林、头孢唑啉、头孢西丁的耐药率分别为98.7％、96.9％、97.5％.结论 阴沟肠杆菌耐药机制复杂,对抗生素具有多重耐药性,临床应合理使用抗菌药物,以减少阴沟肠杆菌耐药性的产生和在院内扩散.     

CTX-M型阴沟肠杆菌插入序列的检测

目的研究插入序列ISEcp1B及超广谱β-内酰胺酶(extended spectrum beta-lactamase,ESBLs)、AmpC酶基因在CTX-M型阴沟肠杆菌(Enterobacter cloacae,ECL)中的分布,并分析其与耐药的关系。方法用MIC法分析2株含CTX-M基因的阴沟肠杆菌的耐药性;用多重聚合酶链反应(PCR)技术扩增ESBLs、AmpC、ISEcp1B基因及I类整合子,对PCR产物进行DNA测序。结果 2株阴沟肠杆菌对青霉素类、四代头孢菌素、喹诺酮类和呋喃类抗生素耐药,对氨基糖苷类和碳青霉烯类表现为敏感。在ECL15同时检出TEM、CTX-M-G1、ACT-1、I类整合子及ISEcp1B基因;在ECL45同时检出SHV、CTX-M-G1及ISEcp1B基因。ECL45(400 bp)ISEcp1B的右端均连接一个反向重复序列,有-35及-10两个启动子,与CTX-M型ESBLs编码起始距离不相同,与ECL15序列一致,GenBank登录号:EF437434;ECL45(500 bp)无反向重复序列,只有一个-35启动子,与CTX-M型ESBLs编码起始距离不相同,与ECL15序列不相同,GenBank登录号:EF441350。结论在CTX-M型阴沟肠杆菌中检出2种新的插入序列ISEcp1B,其对CTX-M的高水平表达和传播可能起重要的调控作用。     

阴沟肠杆菌对β—内酰胺类抗生素耐药的机制

阴沟肠杆菌对β－内酰胺类抗生素日益严重的耐药引起人们的重视,其耐药机制主要有：外膜微孔蛋白的缺失或减少;细菌产生β－内酰胺酶（ｂｌａ）;细菌的青霉素结合蛋白（ＰＢＰｓ）改变导致其对β－内酰胺类抗生素亲和力下降,其中以细菌产生ｂｌａ为最主要的机制。近年来的研究在阴沟肠杆菌Ｉ型ｂｌａ（即可诱导的染色体介导的ｂｌａ）的基因调控方面取得了突破性进展。     

院内感染阴沟肠杆菌的分布及耐药性分析

随着抗生素的广泛使用,带来了严重的细菌耐药性问题。阴沟肠杆菌作为一种条件致病菌可致人体多部位感染,由于其感染率高和耐药率高,越来越引起人们的重视。我们从１９９４年６月～１９９７年６月院内感染的患者的各种标本中分离出３１０株阴沟肠杆菌,测定了对１７种抗...     

164株阴沟肠杆菌的药物敏感性分析

目的了解阴沟肠杆菌的药物敏感性情况以指导临床合理用药。方法对中山大学附属第一医院近3年分离出的阴沟肠杆菌的标本分布及耐药情况进行回顾性分析。结果共分离出164株阴沟肠杆菌,其对亚胺培南、头孢吡肟、阿米卡星和氧氟沙星的敏感性较高。结论阴沟肠杆菌对3代头孢菌素耐药严重,呈多重耐药性,亚胺培南仍为治疗阴沟肠杆菌严重感染的首选用药。     

阴沟肠杆菌临床分离与耐药性的6年监测

目的了解2000年至2005年阴沟肠杆菌在临床标本中的分离情况及其耐药趋势,为临床感染的预防和治疗提供参考资料。方法回顾分析2000年至2005年间所有标本中阴沟肠杆菌的分离率,菌株在标本和病区的分布及对13种抗菌药物的耐药率。结果阴沟肠杆菌的分离率有逐年升高的趋势,从2000年的0.47%增至2005的0.69%;其在呼吸道标本的分离率最高(67.80%),其次是尿液(14.39%)、血液(9.28%);病区以重症监护室最高(29.92%),阴沟肠杆菌对13种抗菌药物的耐药率,2000年仅氨苄西林、妥布霉素、庆大霉素>50%,而到2005年13种抗菌药物除亚胺培南外的耐药率均>50%;其耐药率均呈递增的趋势,以2003年的耐药率最高。耐药率增幅列前3位分别为:头孢他啶33.8%~72.8%、氨曲南41.9%~80.2%、哌拉西林/他唑巴坦48.4%~81.5%。结论阴沟肠杆菌的临床分离率在逐年增加,对多数抗菌药物有较高的耐药性,且耐药率呈整体上升趋势,尤其出现耐亚胺培南的耐药株,应引起重视,合理使用抗菌药物以降低耐药性已经非常重要。     

医院感染阴沟肠杆菌的耐药性及基因型分析

了解中南大学湘雅三医院重症监护病房(ICU)医院感染阴沟肠杆菌的耐药谱及分子流行病学特征,指导临床合理用药。基因分型采用优化反应体系的随机扩增多态性DNA(RAPD)法,耐药分析选用K-B法。RAPD分型将28株阴沟肠杆菌分为11种株型,耐药谱则将其分为12种不同的耐药型。基因分型和耐药分析显示ICU有多重耐药的阴沟肠杆菌株爆发流行现象,它为控制阴沟肠杆菌的医院内感染、追踪传染原、切断传播途径提供遗传学信息,指导临床医生选用敏感有效的抗生素。     

阴沟肠杆菌（nifL—A^c）工程菌株的构建

采用基因定位突变的方法,在体外把来自ｐＢＲ３２２的四环素抗性基因片段插入由ｐＳＴ１１４２质粒所携带的阴沟肠杆菌ｎｉｆＬ基因３－端的ＳｍａⅠ位点、再经细胞体内同源基因片段的重组交换,选择获得了在染色体上ｎｉｆＬ突变的阴不直菌Ｅ１２和Ｅ１３,两者Ｔｃ＾ｒ基因插入ｎｉｆＬ的转录方向相反。经分析显示,Ｅ１３（ｎｉｆＬ）由于Ｔｃ＾ｒ基因插入后,在ｎｉｆＡ上 生具有启动子核苷酸序列（－ＴＴＴＣＡＴＡ－）,激活     

产AmpC酶阴沟肠杆菌的基因分析及其耐药性

探讨昆明地区阴沟肠杆菌的耐药性及与结构基因ampC和调节基因ampD的相关性。通过K-B法检测阴沟肠杆菌的药敏情况,头孢西丁三维试验检测AmpC酶,PCR法扩增ampC和ampD基因。结果显示74株阴沟肠杆菌经头孢西丁三维试验检测,产AmpC酶的有17株,检出率为22.3%,而且产酶菌株抗生素敏感率低于非产酶菌株。ampC基因扩增阳性率为89.2%(66/74);64株ampD基因阳性率为86.5%(64/74)。实验证实昆明地区产酶阴沟肠杆菌耐药状况严重,与结构基因ampC和调节基因ampD密切相关。     

Biodegradation of 2-Chlorobenzoic Acid by Enterobacter cloacae: Growth Kinetics and Effect of Growth Conditions

Enterobacter cloacae was originally isolated from soil irrigated with wastewater on the basis of its ability to grow with linear alkylbenzene sulfonate (LAS) as the sole source for carbon and energy. The isolated bacterium was grown in batch cultures using a 2-chlorobenzoic acid (2-CBA)-containing minimal salt medium (MSM). 2-CBA was found to be the sole source for carbon and energy. 2-CBA inhibited the growth rate with a maximum concentration of 10 mM, after which no growth occurred. The Haldane model was used to predict the specific growth rate concentration data. 2-CBA degradation by starved E. cloaca cells was faster than that of nonstarved cells. The maximum growth rates on 2-CBA (2 mM) for starved and nonstarved cells reached only 0.34 and 0.28 h^?1, respectively. Glucose, lactose, sucrose, maltose, succinic acid, and mannitol as additional carbon sources at a fixed concentration (0.2%) caused the degradation rate of 2-CBA to proceed faster at ranges between 1.08- and 1.5-fold higher than that of the control. In contrast, using only fructose and sorbitol as the carbon sources showed catabolic repression of the degradation activity of 2-CBA by E. cloaca cells, although their cell mass was improved. All nitrogen sources supplied caused an increase in cell mass, whereas only lysine, alanine, glutamine, casein, and yeast extract caused a decrease in the degradation rate of 2-CBA, with a range between 12% and 28%. The activity of C120 could be detected in a crude extract of E. cloacae cells, indicating that the chloroaromatic ring fission occurs through the ortho pathways, not through the meta pathways. The data showed that different initial cell (inocula) densities did not affect the induction time for 2-CBA degradation. However, doubling the initial cell densities reduced the time required for reaching the complete degradation. 2-CBA degradation was optimally achieved at a 37°C incubation temperature and a pH of 7.5.     

Biological and immunological comparisons of Enterobacter cloacae and Escherichia coli porins

Abstract Bacteriocin susceptibilities indicate that during cloacin DF13 uptake the F porin of Enterobacter cloacae plays a similar role to that reported for the OmpF porin of Escherichia coli during colicin A entry. The translocatory activities of these two porins during the bacteriocin uptake can be substituted by the porins D and OmpC, respectively, under conditions not requiring the receptor binding step. Using anti-peptide antibodies, a peptide located in the internal loop L3 of the Escherichia coli OmpF porin was identified in the D and F porins of Enterobacter cloacae. The results demonstrated the existence of a close relationship between porins in terms of both antigenic determinants and bacteriocin susceptibilities.     

利用胞内代谢物变化预测大肠埃希菌耐药性

目的利用胞内代谢物量的差异达到预测大肠埃希氏菌的药物敏感性结果。方法收集临床分离大肠埃希菌120株,在头孢他啶存在条件下培养4 h。收集菌体,测定细胞内化合物然后进行多变量分析。结果 120株大肠埃希菌的聚类分析结果很好地体现了药物敏感性特征,57株耐药菌中,有5株被误判,准确率为91%;47株中敏菌株中,有8株被误判,准确率为83%;16株敏感菌中有2株被误判;准确率为88%。结论该方法能够有效预测菌株的药敏结果,而且快速可靠。     

B-animalisV9质粒检测及抗生素敏感性试验

本实验对动物双歧杆菌Vg（B．animalisvg）菌株进行质粒检测及抗生素敏感性测试。结果表明,Baninalis V9菌株无质粒检出。该菌株对针对G＋菌的抗生素,如青霉素类（青霉素G、氨苄青霉素）和大环内酯类（氯霉素、红霉素）表现出高度敏感;对抑制细菌蛋白质合成的广谱（G＋、G-）抗生素（四环素、利福平、痢特灵）表现出中度敏感;对G-菌的抗生素,如氨基糖苷类（庆大霉素、链霉素）、喹诺酮类（诺氟沙星）及内酰胺类（卡那霉素、丁胺卡那霉素）表现出耐药性;而对广谱抗生素磺胺甲基异嗯唑也表现为耐药。     

Modulation of virulence and antibiotic susceptibility of enteropathogenic Escherichia coli strains by Enterococcus faecium probiotic strain culture fractions

The increasing rate of antimicrobial resistance drastically reduced the efficiency of conventional antibiotics and led to the reconsideration of the interspecies interactions in influencing bacterial virulence and response to therapy. The aim of the study was the investigation of the influence of the soluble and cellular fractions of Enterococcus (E.) faecium CMGB16 probiotic culture on the virulence and antibiotic resistance markers expression in clinical enteropathogenic Escherichia (E.) coli strains.The 7 clinical enteropathogenic E. coli strains, one standard E. coli ATCC 25,922 and one Bacillus (B.) cereus strains were cultivated in nutrient broth, aerobically at 37 °C, for 24 h. The E. faecium CMGB16 probiotic strain was cultivated in anaerobic conditions, at 37 °C in MRS (Man Rogosa Sharpe) broth, and co-cultivated with two pathogenic strains (B. cereus and E. coli O28) culture fractions (supernatant, washed sediment and heat-inactivated culture) for 6 h, at 37 °C. After co-cultivation, the soluble and cellular fractions of the probiotic strain cultivated in the presence of two pathogenic strains were separated by centrifugation (6000 rpm, 10 min), heat-inactivated (15 min, 100 °C) and co-cultivated with the clinical enteropathogenic E. coli strains in McConkey broth, for 24 h, at 37 °C, in order to investigate the influence of the probiotic fractions on the adherence capacity and antibiotic susceptibility. All tested probiotic combinations influenced the adherence pattern of E. coli tested strains. The enteropathogenic E. coli strains susceptibility to aminoglycosides, beta-lactams and quinolones was increased by all probiotic combinations and decreased for amoxicillin-clavulanic acid. This study demonstrates that the plurifactorial anti-infective action of probiotics is also due to the modulation of virulence factors and antibiotic susceptibility expression in E. coli pathogenic strains.     

庆大霉素耐药阴沟肠杆菌16S rRNA甲基化酶基因的研究

目的了解庆大霉素耐药阴沟肠杆菌中16S rRNA甲基化酶基因的分布情况。方法筛选临床分离的庆大霉素耐药的阴沟肠杆菌30株。采用聚合酶链反应（PCR）方法扩增16S rRNA甲基化酶五种相关耐药基因armA、rmtA、rmtB、rmtC和rmtD,PCR产物进行电泳分析和测序,抗菌药物的药物敏感性试验采用纸片扩散法进行,Whonet 5.4软件进行数据分析。结果 30株庆大霉素耐药阴沟肠杆菌中16S rRNA甲基化酶基因的检出率为56.7%（17/30）,其中30.0%（9/30）检出armA基因、26.7%（8/30）检出rmtB基因,未检出rmtA、rmtC和rmtD基因。30株庆大霉素耐药阴沟肠杆菌对妥布霉素、阿米卡星耐药率分别为93.3%,83.3%;对其他抗菌药物除亚胺培南较敏感外,其余抗菌药物耐药率大于60.0%。结论庆大霉素耐药阴沟肠杆菌中16S rRNA甲基化酶基因主要是armA和rmtB基因,碳青霉烯类对该菌有较好体外抗菌活性。     

Kinetics and modeling of hexavalent chromium reduction in Enterobacter cloacae

Kinetics of bacterial reduction of toxic hexavalent chromium (chromate: CrO(4) (2-)) was investigated using batch and fedbatch cultures of Enterobacter cloacae strain HO1. In fedbatch cultures, the CrO(4) (2-) feed was controlled on the basis of the rate of pH change. This control strategy has proven to be useful for avoiding toxic CrO(4) (2-) overload. A simple mathematical model was developed to describe the bacterial process of CrO(4) (2-) reduction. In this model, two types of bacterial cells were considered: induced, CrO(4) (2-)-resistant cells and uninduced, sensitive ones. Only resistant cells were assumed to be able to reduce CrO(4) (2-). These fundamental ideas were supported by the model predictions which well approximated all experimental data. In a simulation study, the model was also used to optimize fed-batch cultures, instead of lengthy and expensive laboratory experiments. (c) 1993 John Wiley & Sons, Inc.